Transcription profiling of Col and CHY-10 mutant in Arabidopsis under cold stress
Ontology highlight
ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
SUBMITTER: Jian-Kang Zhu
PROVIDER: E-MEXP-67 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:In plants, the activation of immunity is often inversely correlated with growth. Mechanisms that plant growth in the context of pathogen challenge and immunity are unclear. Investigating Arabidopsis infection with the powdery mildew fungus, we find that the Arabidopsis atypical E2F DEL1, a transcriptional repressor known to promote cell proliferation, represses accumulation of the hormone salicylic acid (SA), an established regulator of plant immunity. DEL1 deficient plants are more resistant to pathogens and slightly smaller than wild type. The resistance and size phenotypes of DEL1 deficient plants are due to the induction of SA and activation of immunity in the absence of pathogen challenge. Moreover, Enhanced Disease Susceptibility 5 (EDS5), a SA transporter required for elevated SA and immunity, is a direct repressed target of DEL1. Together, these findings indicate that DEL1 control of SA levels contributes to regulating the balance between growth and immunity in developing leaves. Mature, fully expanded leaves of 41/2-week-old Col-0 and del1-1 plants were harvested at 5 days after G. orontii infection and from uninfected control plants for RNA extraction and hybridization on Affymetrix microarrays.
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Project description:Obligate biotrophs such as the virulent powdery mildew Golovinomyces orontii alter plant host cellular architecture, metabolism, and defense in order to acquire nutrients while suppressing cell death and senescence. G. orontii exclusively infects epidermal cells of Arabidopsis with clearly defined stages of infection. Host factors mediating the powdery mildew (PM) interaction are often expressed in the mesophyll cells underlying the infected epidermal cells. Therefore, in order to identify Arabidopsis processes and regulators mediating this PM interaction, we used UV laser microdissection to isolate cells at the PM infection site for global expression profiling. As part of this process, we optimized and/or developed novel tissue preparation, RNA extraction and amplification, and quality control protocols resulting in highly correlated biological replicate data. We focused on the growth and reproduction stage of the PM infection (5 days post infection) when the number of reproductive structures, conidiophores, can be quantified. Site-specific profiling increased our sensitivity dramatically, allowing us to identify specific processes, process components, and their putative regulators hidden in previous whole leaf global expression analyses. For example, the known cell cycle regulator MYB3R4 exhibits altered expression at the site of infection, as do a subset of cell-cycle-associated genes. Furthermore, null myb3r4 mutants exhibit enhanced resistance to PM with reduced conidiophores per colony, suggesting cell cycle control plays an important role in the PM interaction. Experiment Overall Design: Arabidopsis whole leaves from wild type Columbia-0 and enhanced disease susceptibility mutant eds16-1, a null isochorismate synthase 1 (At1g74710) mutant were harvested at 5 days after Golovinomyces orontii infection, microwave-fixed, paraffin-embedded and sectioned. Groups of epidermal and mesophyll cells (~20 cells/group) surrounding the G. orontii infected epidermal cell were cut using a Leica AS laser microdissection (LMD) system. In parallel, groups of epidermal and mesophyll cells were collected from uninfected leaves at 5 days from wild type Arabidopsis. LMD-isolated cells, whole leaf, whole leaf amplified and tissue scrape samples were used for RNA extraction and hybridization to Affymetrix Arabidopsis ATH1 microarrays. Gene expression profiles were obtained for wild type from all samples and for ics1 mutant from LMD infected samples. The experiment includes 2 biological replicates.
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Project description:RNA-seq reads from the selfing species Arabidopsis thaliana were produced from flowers to study the consequences of the transition from the ancestral state (outcrossing) to the derived state (selfing). This was done in the context of examining another species in the Arabidopsis genus (A. lyrata) and another species pair (Capsella rubella versus Capsella grandiflora, which are selfing and outcrossing, respectively). These samples were generated to complement part of this larger study. Briefly, the shift from outcrossing to selfing is common in flowering plants, but neither the genomic consequences nor the speed with which they appear are well understood. An excellent model for understanding the evolution of self fertilization is provided by Capsella rubella, which became self-compatible <200,000 years ago. We present a reference genome for the species, and compare RNA expression and polymorphism patterns between C. rubella and its outcrossing progenitor C. grandiflora. There is a clear shift in the expression of genes associated with flowering phenotypes; a similar shift is seen in the related genus Arabidopsis, where self-fertilization evolved about 1 million years ago. DNA sequence polymorphisms distinguishing the two Capsella species reveal rapid genome-wide relaxation of purifying selection in C. rubella but without a concomitant change in transposable element abundance. Overall, we document that the transition to selfing may be typified by shifts in expression for genes that function in pollen and flower development, along with a measurable reduction of purifying selection. As part of a cross-species comparison of gene expression, RNA-seq data was generated in biological replication (2 replicates) from Arabidopsis thaliana at the floral stage. In total, two samples (biological replicates) were used. The reference strain was used for the experments (strain Col-0). Resulting data about gene expression was used as part of a larger study. The Capsella rubella and Capsella grandiflora data are included in GEO Series GSE45518.
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